exchange reactions catalyzed by methioninase from pseudomonas putida.highly purified methioninase from pseudomonas putida, which catalyzes alpha, gamma-elimination reactions of homocysteine and its s-substituted derivatives as well as alpha, beta-elimination reactions of cysteine and its derivatives, was found to catalyze exchange reactions between the substituent at the gamma-carbon of homocysteine substrates and exogenously added alkanethiols, forming the corresponding s-alkylhomocysteines. it also catalyzed similar beta-exchange reactions between cysteine and ...19751213994
dynamic and steady state studies of phenol biodegradation in pure and mixed cultures.the microbial degradation of phenol by pure and mixed cultures of pseudomonas putida was studied in batch, phenol-stat, and continuous culture systems. in the continuous culture runs, both steady state and transient experiments were performed. from these experiments, a model for the kinetic behavior of the organisms was evolved and an analysis performed on the stability and dynamic behavior of pure and mixed cultures. the results indicate that it should be possible to achieve phenol removal from ...19751236402
[r factor found in pseudomonas putida]. 19751240230
role and regulation of the ortho and meta pathways of catechol metabolism in pseudomonads metabolizing naphthalene and salicylate.the enzymes of naphthalene metabolism are induced in pseudomonas putida atcc 17484, ppg7, ncib 9816, and pg and in pseudomonas sp. atcc 17483 during growth on naphthalene or salicylate; 2-aminobenzoate is a gratuitous inducer of these enzymes. the meta-pathway enzymes of catechol metabolism are induced in atcc 17483 and ppg7 during growth on naphthalene or salicylate or during growth in the presence of 2-aminobenzoate, but in atcc 17484 and ncib 9816 the ortho-pathway enzymes of catechol metabol ...19761245462
properties of an inducible uptake system for beta-ketoadipate in pseudomonas putida.wild-type strains of pseudomonas putida form an inducible uptake system that appears to act on beta-ketoadipate under normal physiological conditions. the system is induced by beta-ketoadipate and is represented by catabolites derived from it. adipate is metabolized very slowly by wild-type p. putida cultures; [14c]adipate was used as an analogue of beta-ketoadipate to measure the transport activity in wild-type cells and in cells that constitutively produced the uptake system. constitutive cell ...19761245464
purification, crystallization and properties of iron-containing superoxide dismutase from pseudomonas ovalis.three electrophoretically distinct superoxide dismutases (ec were observed in the crude extracts from pseudomonas ovalis. one of these was isolated as an iron-containing superoxide dismutase. it contained 1.4 gatoms of fe per mol of enzyme, and had a specific activity of 3900 units per mg of protein. a crystallized enzyme contained 1.1 gatoms of fe per mol of enzyme, and had a specific activity of 3100 units per mg of protein. the results of sedimentation equilibrium and gel filtration ...19761247598
ubiquity of plasmids in coding for toluene and xylene metabolism in soil bacteria: evidence for the existence of new tol plasmids.thirteen bacteria have been isolated from nine different soil samples by selective enrichment culture on m-toluate (m-methylbenzoate) minimal medium. eight of these were classified as pseudomonas putida, one as a fluorescent pseudomonas sp., and four as nonfluorescent pseudomonas sp. all 13 strains appeared to carry tol plasmids superficially similar to that previously described in p. putida mt-2 in that: (i) all the wild-type strains could utilize toluene, m-xylene, and p-xylene as sole carbon ...19761254555
metabolism of resorcinylic compounds by bacteria: orcinol pathway in pseudomonas putida.enrichment cultures yielded two strains of pseudomonas putida capable of growth with orcinol (3,5-dihydroxytoluene) as the sole source of carbon. experiments with cell suspensions and cell extracts indicate that orcinol is metabolized by hydroxylation of the benzene ring followed successively by ring cleavage and hydrolyses to give 2 mol of acetate and 1 mol of pyruvate per mol of orcinol as shown: orcinol leads to 2,3,5-trihydroxytoluene leads to 2,4,6-trioxoheptanoate leads to acetate + acetyl ...19761254564
peptide utilization by pseudomonas putida and pseudomonas maltophilia.pseudomonas putida assimilates peptides and hydrolyses them with intracellular peptidases. amino acid auxotrophs (his, trp, thr or met) grew on a variety of di- and tripeptides up to twice as slowly as with free amino acids. pseudomonas putida has separate uptake systems for both dipeptides and oligopeptides (three or more residues). although the dipeptide system transported a variety of structurally diverse dipeptides it did not transport peptides having either unprotonatable n-terminal amino g ...19761255131
intracellular peptide hydrolysis by pseudomonas putida and pseudomonas maltophilia.amino acids liberated by peptidase hydrolysis of di- and oligopeptides by pseudomonas putida were measured by trinitrobenzenesulphonate assay and high voltage electrophoresis or paper chromatography followed by ninhydrin spray. intact bacteria or periplasmic contents released by lysozyme treatment did not hydrolyse peptides. subcellular fractionation showed that glycylmethionine peptidase activity was cytoplasmic. this enzyme had a km of 2 mm, and was stimulated fivefold by i mm-co2+. crude pept ...19761255132
constitutive synthesis of enzymes of the protocatechuate pathway and of the beta-ketoadipate uptake system in mutant strains of pseudomonas putida.mutant pseudomonas putida strains that produce constitutive levels of the beta-ketoadipate uptake system are selected by the sequential transfer of cultures between mineral growth media supplemented with the noninducing growth substrate succinate and growth media containing beta-ketoadipate as the sole carbon and energy source. the mutant strains also produce constitutively three catabolic enzymes that give rise to beta-ketoadipate from the metabolic precursor beta-carboxy-cis, cis-muconate, and ...19761262305
evidence for autogenous regulation of pseudomonas putida tryptophan synthase.studies of a trpa mutant constitutive for tryptophan synthase production support the hypothesis of autogenous regulation (r. f. goldberger, 1974; a. r. proctor and i. p. crawford, 1975) of the pseudomonas putida trpab loci.19761262309
the hydroxylation of p-cresol and its conversion to p-hydroxybenzaldehyde in pseudomonas putida. 19761267796
supra-operonic clustering of genes specifying glucose dissimilation in pseudomonas putida.the linkage arrangements of genes governing glucolysis in pseudomonas putida have been determined by transductional analysis. five genes (gdh, kgta, kgtb, edd and eda), comprising at least three operons, are contransducible with each other, but not with ggu (glucose and gluconate uptake) nor with genes of a known supra-operonic cluster of genes specifying enzymes of other dissimilatory pathways, nor with a biochemically uncharacterized his marker. it thus appears that p. putida may have more tha ...19761272248
[comparative study of naphthalene catabolism routes in 2 strains of pseudomonas putida]. 19761277993
bacterioplankton community structure and dynamics after large-scale release of nonindigenous bacteria as revealed by low-molecular-weight-rna analysis.a set of freshwater mesocosms (1.7 m3 each) was inoculated with large amounts of escherichia coli, pseudomonas putida, and their culture medium to substantially disturb the natural microbial community. to monitor microbial community dynamics, low-molecular-weight rna (5s rrna and trna) obtained directly from bacterioplankton was analyzed by using high-resolution electrophoresis. the introduced bacteria showed no significant effect on the community structure of the natural bacterial assemblage an ...19921280060
in vitro interactions of pseudomonas rna polymerases with tac and rna i promoters.the activities of rna polymerases from pseudomonas putida and pseudomonas aeruginosa were compared with that of escherichia coli rna polymerase in an in vitro transcription system. all three enzymes initiated transcription at the tac promoter and the rna i promoter of e. coli. we measured the rate of open complex formation between the rna polymerases and the promoters, and the saturation level of open complex formation at equilibrium in single-round transcription. the relative rates of open comp ...19921282050
[use of bacterial toximeters with separate cell cultures for continuous water monitoring].two commercially available bacterial toxicity monitors are compared. as test organisms pure cultures of pseudomonas putida are used in both systems. the bacteria are grown continuously in turbidostatic ("toxalarm", lar, berlin, germany) or chemostatic ("stiptox-norm", siepmann und teutscher, gross-umstadt, germany) regulated cultures in a selective culture medium in nonsterile devices. toxic substances can be detected by continuously working bacterial respiration tests. oxygen consumption is the ...19921284920
[role of phenylalanine in the biosynthesis of fluorescent pigment in pseudomonas putida bacteria].the contemporary data of the participation of phenylalanine in the biosynthesis of fluorescent pigment pyoverdine pm of pseudomonas putida strain m are presented. using aro1phu1 mutant of this strain, it has been shown that one of the precursors of the dihydroxyquinoline moiety of the pyoverdine pm is phenylalanine in the d- or l-form. these results were confirmed in experiments with 14c-phenylalanine incorporation. pyoverdine pm that was synthesized by aro1phu1 mutant from exogenous phenylalani ...19921287406
[regulation of the expression of plasmid determination responsible for caprolactam degradation by bacteria of the genus pseudomonas].on the basis of the study of some tn5 induced mutants in pseudomonas putida strain bs836 containing the plasmid pbs268 coding caprolactam degradation, growth on caprolactam and its intermediates, and the data on the induction of oxidative activities in plasmid containing p. putida strain bs831 it was shown that plasmid and chromosome genes regulated the expression of cap-determinants. the regulation has some elements of the negative control mechanism. caprolactam is the inducer of the synthesis ...19921287407
[purification and properties of pyrocatechase ii from pseudomonas putida strain 87].induction of modified ortho-pathway enzymes (catechol 1.2-dioxygenase ii, muconate cycloisomerase ii, dienelactone hydrolase, and maleylacetate reductase) was found in pseudomonas putida 87, when 3-chlorobenzoic acid was used as a sole carbon and energy source. catechol 1.2-dioxygenase ii, the key chlorocatechol cleaving enzyme, was purified and characterized. the enzyme molecular mass as determined by gel filtration was 65,000 da; the minimum molecular mass upon sds electrophoresis was 33,000 d ...19921294257
[preservation of pseudomonas putida bacteria at low temperatures].we have found that the mode of cooling, composition of cryopreservation medium, original concentration of cells and storage temperature affect viability of pseudomonas putida bacteria during low-temperature preservation. we have elaborated the conditions of preservation, providing for a high survival of bacteria, namely: one-stage cooling with the rates of 30, 40 degrees c/min or immersion into liquid nitrogen in the culturing medium with addition of sucrose, glycerol or dimexide in the concentr ...19921297044
[plasmid p85 from azospirillum brasilense sp245: study of the circle of possible hosts and incompatibility with plasmids from azospirillum brasilense sp7].the possibility of the stable inheritance of the plasmid p85 mobilized derivatives from azospirillum brasilense sp245 in the cells of the bacterial genera rizobiaceae (agrobacterium tumfaciens) and pseudomonadaceae (pseudomonas putida) has been shown. the plasmid p85 participates in coding for the physiologically active products (the plant hormones). it is not inherited by the escherichia coli strains. for the first time the incompatibility of azospirillium plasmids has been demonstrated on the ...19921298886
[cloning and expression of the arthrobacter globiformis fcb genes in bacillus subtilis].the fcb genes of arthrobacter globiformis kzt1 coding for the dehalogenase (4-chlorobenzoate-4-hydroxylase) activity have been cloned. the characteristics of fcb genes expression have been studied. the recombinant strains of bacillus subtilis 6jm15 (pcbs 311) and 6jm15 (pcbs1) have shown the decreased level of substrate dehalogenation as compared with the one in the parent strain kzt1 and the recombinant strains of escherichia coli and pseudomonas putida.19921301499
iron uptake and molecular recognition in pseudomonas putida: receptor mapping with ferrichrome and its biomimetic analogs.the presence of an fe(3+)-ferrichrome uptake system in fluorescent pseudomonas spp. was demonstrated, and its structural requirements were mapped in pseudomonas putida with the help of biomimetic ferrichrome analogs. growth tests, 55fe3+ uptake, and competition experiments demonstrated that the synthetic l-alanine derivative b5 inhibits the action of ferrichrome but does not facilitate fe3+ transport, while the enantiomeric d-ala derivative b6 fails to compete with ferrichrome. contraction of th ...19921309523
the dehalogenase gene dehi from pseudomonas putida pp3 is carried on an unusual mobile genetic element designated a result of the production of two dehalogenases (dehi and dehii), pseudomonas putida pp3 utilized halogenated alkanoic acids, such as 2-monochloropropionic acid (2mcpa), as sole sources of carbon and energy. the dehi gene (dehi) was carried on a mobile genetic element (deh) located on the chromosome of strain pp3. deh recombined with target plasmid dnas at high frequencies (e.g. 3.8 x 10(-4) per rp4.5 plasmid transferred). the regulated expression of dehi was detected in p. putida, pseudomona ...19921312533
localization and functional analysis of structural and regulatory dehalogenase genes carried on deh from pseudomonas putida pp3.pseudomonas putida pp3 expressed two dehalogenases, dehi and dehii. the dehi gene (dehi) was located on a mobile dna element (deh) which inserted at high frequencies into target plasmids from its chromosomal location. from a recombinant tol plasmid (pww0) containing a 6.0-kb deh element inserted into the plasmid's 5.6-kb ecori-g restriction endonuclease fragment, an 11.6-kb ecori fragment was cloned. subcloning analysis and insertion mutagenesis produced a structural map of the deh element and l ...19921312534
nmr studies of azotobacter vinelandii and pseudomonas putida seven-iron ferredoxins. direct assignment of beta-cysteinyl carbon nmr resonances and further proton nmr assignments of cysteinyl and aromatic resonances.ferredoxins are proteins which contain iron and inorganic sulfide and are capable of electron transport. they are found in a wide range of organisms, from anaerobic bacteria, to plants and mammals. although nmr spectroscopy has been used to study ferredoxins since the 1970s, little important structural or biochemical information has resulted from these investigations. the major difficulty has been the effect of the paramagnetic iron-sulfur clusters on the peptide resonances, hindering nuclear ov ...19921314816
cloning, sequencing, and overexpression of a [2fe-2s] ferredoxin gene from escherichia coli.escherichia coli contains a soluble, [2fe-2s] ferredoxin of unknown function (knoell, h.-e., and knappe, j. (1974) eur. j. biochem. 50, 245-252). using antiserum to the purified protein to screen e. coli genomic expression libraries, we have cloned a gene (designated fdx) encoding this protein. the dna sequence of the gene predicts a polypeptide of 110 residues after removal of the initiator methionine (polypeptide m(r) = 12,186, holoprotein m(r) = 12,358). the deduced amino acid sequence is str ...19921317854
a general system to integrate lacz fusions into the chromosomes of gram-negative eubacteria: regulation of the pm promoter of the tol plasmid studied with all controlling elements in monocopy.a new procedure is described to recombine plasmid-borne lacz fusions into the chromosome of gram-negative eubacteria in order to study promoter activity in monocopy. the procedure is based upon the insertion into the chromosome of a target bacterium of a recombinant transposon that carries dna sequence homology to the regions flanking lacz fusions present in multicopy promotor-probe vectors, which can be mobilized via rp4-mediated transfer but are unable to replicate in non-enteric bacteria. dou ...19921318499
isolation and molecular characterization of a novel broad-host-range plasmid from bordetella bronchiseptica with sequence similarities to plasmids from gram-positive organisms.a 2.6 kb plasmid, named pbbr1, was isolated from bordetella bronchiseptica s87. after insertion of an antibiotic resistance marker, this plasmid could be transferred into escherichia coli, bordetella pertussis, b. bronchiseptica, vibrio cholerae, rhizobium meliloti, and pseudomonas putida by transformation or conjugation. conjugation was possible only when the incp group transfer functions were provided in trans. as shown by incompatibility testing, pbbr1 does not belong to the broad-host-range ...19921321324
molecular cloning and mapping of phenol degradation genes from bacillus stearothermophilus fdtp-3 and their expression in escherichia coli.two genes of the meta pathway of phenol degradation were cloned from a phenol-utilizing strain of bacillus stearothermophilus and were mapped by subcloning and by use of a tn5 insertion mutation. they code for phenol hydroxylase and catechol 2,3-dioxygenase, respectively. the gene encoding catechol 2,3-dioxygenase, which is more thermostable than catechol 2,3-dioxygenase encoded by the other gene, shares rather limited homology with that from pseudomonas putida.19921325141
the refined crystal structure of pseudomonas putida lipoamide dehydrogenase complexed with nad+ at 2.45 a resolution.the three-dimensional structure of one of the three lipoamide dehydrogenases occurring in pseudomonas putida, lipdh val, has been determined at 2.45 a resolution. the orthorhombic crystals, grown in the presence of 20 mm nad+, contain 458 residues per asymmetric unit. a crystallographic 2-fold axis generates the dimer which is observed in solution. the final crystallographic r-factor is 21.8% for 18,216 unique reflections and a model consisting of 3,452 protein atoms, 189 solvent molecules and 4 ...19921325638
gonococcal transferrin-binding protein 1 is required for transferrin utilization and is homologous to tonb-dependent outer membrane receptors.the pathogenic neisseria species are capable of utilizing transferrin as their sole source of iron. a neisserial transferrin receptor has been identified and its characteristics defined; however, the biochemical identities of proteins which are required for transferrin receptor function have not yet been determined. we identified two iron-repressible transferrin-binding proteins in neisseria gonorrhoeae, tbp1 and tbp2. two approaches were taken to clone genes required for gonococcal transferrin ...19921325963
purification and characterisation of the nadh:acceptor reductase component of xylene monooxygenase encoded by the tol plasmid pww0 of pseudomonas putida mt-2.the xylene monooxygenase system encoded by the tol plasmid pww0 of pseudomonas putida catalyses the hydroxylation of a methyl side-chain of toluene and xylenes. genetic studies have suggested that this monooxygenase consists of two different proteins, products of the xyla and xylm genes, which function as an electron-transfer protein and a terminal hydroxylase, respectively. in this study, the electron-transfer component of xylene monooxygenase, the product of xyla, was purified to homogeneity. ...19921327782
cloning and transposon vectors derived from satellite bacteriophage p4 for genetic manipulation of pseudomonas and other gram-negative bacteria.we developed transposon and cloning shuttle vectors for genetic manipulation of pseudomonas and other gram-negative bacteria, exploiting the unique properties and the broad host range of the satellite bacteriophage p4. p4::tn5 ap-1 and p4::tn5 ap-2 are suicide transposon vectors which have been used for efficient tn5 mutagenesis in pseudomonas putida. pkgb2 is a phasmid vector with a cloning capacity of about 7.5 kb; useful unique cloning sites are saci and sacii in the streptomycin resistance d ...19921329125
environmentally directed mutations in the dehalogenase system of pseudomonas putida strain pp3.favourable mutations involving the two dehalogenases (dehi and dehii) of pseudomonas putida pp3 and derivative strains containing the cloned gene for dehi (dehi) occurred in response to specific environmental conditions, namely: starvation conditions; the presence of dehalogenase substrates (halogenated alkanoic acids--haas) which were toxic to p. putida; and/or the presence of a potential growth substrate. fluctuation tests showed that these mutations were environmentally directed by the presen ...19921332636
4-oxalocrotonate tautomerase, an enzyme composed of 62 amino acid residues per monomer.the xylh gene encoding 4-oxalocrotonate tautomerase (4-ot) has been located on a subclone of the pseudomonas putida mt-2 tol plasmid pww0 and inserted into an escherichia coli expression vector. several of the genes of the metafission pathway encoded by pww0 have been cloned in e. coli, but the overexpression of their gene products has met with limited success. by utilizing the e. coli alkaline phosphatase promoter (phoa) coupled with the proper positioning of a ribosome-binding region, we are a ...19921339435
bioluminescence-based detection of genetically engineered microorganisms in nonsterile river water.the luminescence genes of the marine bacterium vibrio fischeri were cloned into a lac expression vector and introduced into escherichia coli and pseudomonas putida. survival of the cells in river water samples was monitored by light measurements. whereas e. coli survived in sterilized river water for more than 29 days, it died off in nonsterile river water after 9 to 13 days. the engineered p. putida cells survived in nonsterile river water for more than 137 days. the detection limit for e. coli ...19921341987
the effect of microcosm design on the survival of recombinant pseudomonas putida in lake water.the survival of pseudomonas putida marked with the xyle gene was monitored in lake-water microcosms. various designs of microcosms were compared. these ranged from 250-ml conical flasks containing 100 ml surface lake water to 12-1 glass containers with lake water overlying sediment, continuous aeration and a supply of fresh surface lake water. the presence of a low flow-through rate was shown to have little effect on the survival of p. putida. an increase in the size of microcosm, presence of se ...19921342636
biodegradation of phenoxyacetic acid in soil by pseudomonas putida pp0301(pr0103), a constitutive degrader of 2,4-dichlorophenoxyacetate.the efficacy of using genetically engineered microbes (gems) to degrade recalcitrant environmental toxicants was demonstrated by the application of pseudomonas putida pp0301(pr0103) to an oregon agricultural soil amended with 500 micrograms/g of a model xenobiotic, phenoxyacetic acid (paa). p. putida pp0301(pr0103) is a constitutive degrader of 2,4-dichlorophenoxyacetate (2,4-d) and is also active on the non-inducing substrate, paa. paa is the parental compound of 2,4-dichlorophenoxyacetic acid ...19921344988
use of a novel plasmid to monitor the fate of a genetically engineered pseudomonas putida strain.plasmid psi30 was constructed to increase the sensitivity of detection of a genetically engineered micro-organism (gem) and its recombinant dna in environmental samples. this broad host-range, mobilizable plasmid contained chlorocatechol (clc) degradative genes, antibiotic resistance genes (ampicillin and kanamycin) and a fragment of eukaryotic dna. the clc genes encode enzymes that convert 3-chlorocatechol to maleylacetic acid permitting the host, pseudomonas putida rc-4, to grow on 3-chloroben ...19921344990
effects of gold(i) antiarthritic drugs and related compounds on pseudomonas putida.the effects of the antiarthritic drugs aurothiomalate (autm), aurothioglucose (autg), auranofin, its metabolite triethylphosphinegold(i)thioglucose (et3pautg), and several related complexes on the growth of pseudomonas putida were studied. two strains were used, one of which (bk135) was more sensitive to et3pautg (tolerant up to 4 microm) than the other (bk403; tolerant to at least 500 microm). gold thiolate complexes and thiolate ligands alone had little effect on growth. gold phosphine complex ...19921355789
isolation, sequencing and expression in e. coli of the urocanase gene from white clover (trifolium repens).the urocanase gene was detected in a clone obtained from a genomic library of white clover. the entire gene has been sequenced and expressed in the pt7-7/e. coli bl 21 (de 3) system. the deduced sequence of the plant urocanase is 72% homologous with that of the well-characterized urocanase from pseudomonas putida. the purification procedure, as well as kinetic and electrophoretic behaviour, of the new enzyme are described.19921356832
dna fingerprints of pseudomonas spp. using rotating field electrophoresis.rotating field electrophoresis (rfe) was applied to evaluate the usefulness of this technique for identification of several pseudomonas strains with suspected importance in deliberate releases. genomes of common wild-type or genetically modified strains of pseudomonas fluorescens, pseudomonas stutzeri, pseudomonas putida and pseudomonas aeruginosa were digested with rare-cutting restriction endonucleases and subjected to pulsed field gel electrophoresis. restrictions with spei or xbai produced 1 ...19921364137
molecular cloning of the xyll-xyle region from the p. putida tol plasmid, pdk1.a 5.2 kilobase ecori restriction fragment from the pseudomonas putida hs1 tol plasmid pdk1, encoding a portion of the lower toluene degradation pathway, was cloned into the e. coli plasmid pbr325. a detailed map of the restriction endonuclease sites was constructed and the nucleotide sequence of three contiguous xhoi fragments, with a combined total length of approximately 3.9 kilobases, has been investigated. this region was determined to contain a total of four separate open reading frames, ea ...19901366507
mechanism of formaldehyde biodegradation by pseudomonas putida.formaldehyde biodegradation by a strain of pseudomonas putida has been studied. the results indicate that this biodegradation is initiated by a dismutation reaction, yielding as products formic acid and methanol. the degradation of methanol and formic acid begins after exhaustion of formaldehyde in the medium, and presents a diauxic pattern: first formic acid is consumed followed by methanol. moreover, cell viability, which is affected by the amount of added formaldehyde, has been determined.19901366532
immobilized and free cell continuous cultures of a recombinant e. coli producing catechol 2,3-dioxygenase in a two-stage chemostat: improvement of plasmid stability.the immobilization of recombinant strains of e. coli w3110/ptg205 in k-carrageenan gel beads improves the plasmid stability during continuous cultures in the absence of selection pressure. since, xyl e gene (which encodes catechol 2,3-dioxygenase from pseudomonas putida) transcription is controlled by the trp promoter, the effects of tryptophan (repressor) and 3 beta-indolyl acrylic acid (derepressor) on ptg 205 stability and enzyme production have been studied in both free and immobilized cell ...19901366935
broad host-range vector for efficient expression of foreign genes in gram-negative bacteria.a broad host-range expression plasmid was constructed comprising the incq replicon, the reca promoter from escherichia coli and the g10-l ribosome binding site (rbs) derived from bacteriophage t7. the structural genes for porcine somatotropin (pst) and e. coli beta-galactosidase (lacz) were used to monitor gene expression in a diverse collection of gram-negative bacterial hosts: escherichia coli, pseudomonas aeruginosa, pseudomonas syringae, pseudomonas putida, pseudomonas fluorescens, pseudomon ...19911367537
degradation of phenol and phenolic compounds by pseudomonas putida ekii.the phenol-degrading strain pseudomonas putida ekii was isolated from a soil enrichment culture and utilized phenol up to 10.6 mm (1.0 g.l-1) as the sole source of carbon and energy. furthermore, cresols, chlorophenols, 3,4-dimethylphenol, and 4-chloro-m-cresol were metabolized as sole substrates by phenol-grown resting cells of strain ekii. under conditions of cell growth, degradation of these xenobiotics was achieved only in co-metabolism with phenol. phenol hydroxylase activity was detectable ...19921368244
maintenance and killing efficiency of conditional lethal constructs in pseudomonas putida.conditional lethal (suicidal) genetic constructs were designed and employed in strains of pseudomonads as models for containment of genetically-engineered microbes that may be deliberately released into the environment. a strain of pseudomonas putida was formed with a suicide vector designated pbap24h that was constructed by cloning the host killing gene (hok) into the rsf1010 plasmid pvdtac24 and placing it under the control of the tac promoter. after hok induction in p. putida only 40% of surv ...19921368479
purification and characterization of a dna-dependent rna polymerase from pseudomonas putida.dna-dependent rna polymerase (ec was purified from pseudomonas putida. the enzyme had the typical composition of beta',beta,alpha, and sigma subunits of eubacterial rna polymerases. the molecular masses of the subunits were 156,000 da, 151,000 da, 87,000 da, and 42,000 da, respectively, as measured by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. the nh2-terminal amino acid residues of the alpha subunit had a marked homology with those of the alpha subuni ...19921369075
monitoring a genetically engineered bacterium in a freshwater environment by rapid enzymatic amplification of a synthetic dna "number-plate".in order to set up a sensitive and reliable detection method to monitor environmentally released genetically engineered microorganisms (gems) a 72-bp, double-stranded dna fragment has been built by annealing and ligating four synthetic oligonucleotides. binding sites for two 20-mer oligonucleotides are situated inside the dna fragment, flanking the centre. into the central part of the construction a 30-nucleotide identification sequence has been fitted. thanks to the presence of the two oligonuc ...19911369367
tracking genetically engineered bacteria: monoclonal antibodies against surface determinants of the soil bacterium pseudomonas putida 2440.assessment of potential risks involved in the release of genetically engineered microorganisms is facilitated by the availability of monoclonal antibodies (mabs), a tool potentially able to monitor specific organisms. we raised a bank of mabs against the soil bacterium pseudomonas putida 2440, which is a host for modified tol plasmids and other recombinant plasmids. three mabs, 7.3b, 7.4d, and 7.5d, were highly specific and recognized only p. putida bacteria. furthermore, we developed a semiquan ...19921373718
molecular biology of the 2-haloacid halidohydrolase iva from pseudomonas cepacia mba4.the structural gene (hdl iva) for the pseudomonas cepacia mba4 2-haloacid halidohydrolase iva (hdl iva) was isolated on a 1.6 kb fragment of ps. cepacia mba4 chromosomal dna. the recombinant halidohydrolase was expressed in escherichia coli and pseudomonas putida and the structural gene was subcloned on to the tac expression vector pbtac1. high-level expression from the tac promoter was seen to be temperature-dependent, a consequence of the nucleotide sequence adjacent to the fragment encoding t ...19921376111
[pyoverdins from pseudomonas putida].the structures of two pyoverdins (pp1 and pp2) and one dihydropyoverdin (dihydro-pp2) from a strain of pseudomonas putida have been elucidated by spectroscopic methods and degradation studies. the pyoverdins pp1 and pp2 consist of a chromophore which was identified as (1s)-5-amino-2,3-dihydro-8,9-dihydroxy-1 h-pyrimido[1,2-a]quinoline-1- carboxylic acid substituted at the amino group with a 3-carboxypropanoyl or a succinamoyl residue and at the carboxy group with the n-terminus of l-ser-l-thr-d- ...19921388514
3-carboxy-cis,cis-muconate lactonizing enzyme from pseudomonas putida is homologous to the class ii fumarase family: a new reaction in the evolution of a mechanistic motif.the gene (pcab) for 3-carboxymuconate lactonizing enzyme (cmle; 3-carboxymuconate cycloisomerase; ec from pseudomonas putida has been cloned into pmg27ns, a temperature-sensitive expression vector, and expressed in escherichia coli n4830. the specific activity and kinetic parameters of the recombinant cmle were comparable to those previously reported. a comparison of the deduced amino acid sequence of cmle with sequences available in the pir and genbank databases revealed that cmle has ...19921390752
purification and characterization of a novel enzyme, arylalkyl acylamidase, from pseudomonas putida sc2.a novel enzyme, arylalkyl acylamidase, which shows a strict specificity for n-acetyl arylalkylamines, but not acetanilide derivatives, was purified from the culture broth of pseudomonas putida sc2. the purified enzyme appeared to be homogeneous, as judged by native and sds/page. the enzyme has a molecular mass of approximately 150 kda and consists of four identical subunits. the purified enzyme catalyzed the hydrolysis of n-acetyl-2-phenylethylamine to 2-phenylethylamine and acetic acid at the r ...19921396711
performance of an aquatic multispecies system in evaluating the effects of a model microbial pest control agent on nontarget organisms.a recirculating multispecies test system was developed in conjunction with a study of the fate and persistence of a model microbial pest control agent on non-target marine and freshwater organisms. the basic unit of the system was a 113-i glass aquarium with vertical biological filters in the center of the aquarium, such that two compartments were formed. this allowed the sequestration of predator and prey species within the same system. organisms from six phyletic groups were subjected to a gen ...19921404484
[the effect of physicochemical factors on the growth of pseudomonas putida bs-2 on a medium with diethylene glycol].the physicochemical factors of medium have been studied for their effect on the physiological indices of growth of pseudomonas putida bs-2 culture utilizing diethylenglycol as the only source of carbon. action of the supraoptimal temperature on the growth process of p. putida bs-2 is accompanied by a decrease (more than twice) in economic coefficient of substrate and specific growth rate as compared with their maximal values. dependences of specific growth rate of p. putida bs-2 in the medium wi ...19921406384
identification of a new gene, tmof, in the pseudomonas mendocina kr1 gene cluster encoding toluene-4-monooxygenase.five genes, tmoabcde, encoding toluene-4-monooxygenase (t4mo) were previously mapped to a 3.6-kb region of a 10.2-kb saci dna fragment isolated from pseudomonas mendocina kr1 (k.-m. yen, m. r. karl, l. m. blatt, m. j. simon, r. b. winter, p. r. fausset, h. s. lu, a. a. harcourt, and k. k. chen, j. bacteriol. 173:5315-5327, 1991). in this report, we describe the identification and characterization of a dna region in the saci fragment whose expression enhances the t4mo activity determined by the t ...19921429451
functional domains of aromatase cytochrome p450 inferred from comparative analyses of amino acid sequences and substantiated by site-directed mutagenesis experiments.several functional domains, especially the active site regions, in aromatase cytochrome p450 were inferred by alignment of amino acid sequences of the enzyme from five species, human, rat, mouse, chicken, and trout, and that of pseudomonas putida cytochrome p450cam, whose x-ray structure has been determined (poulos, t.l., finzel, b.c., and howard, a.j. (1987) j. mol. biol. 195, 687-700). the predicted functions of these domains have been evaluated by site-directed mutagenesis. eighteen mutants, ...19921429608
expression and transfer of engineered catabolic pathways harbored by pseudomonas spp. introduced into activated sludge microcosms.two genetically engineered microorganisms (gems), pseudomonas sp. strain b13 fr1(pfrc20p) (fr120) and pseudomonas putida kt2440(pwwo-eb62) (eb62), were introduced into activated sludge microcosms that had the level of aeration, nutrient makeup, and microbial community structure of activated sludge reactors. fr120 contains an experimentally assembled ortho cleavage route for simultaneous degradation of 3-chlorobenzoate (3cb) and 4-methyl benzoate (4mb); eb62 contains a derivative tol plasmid-enco ...19921444370
stereospecific hydroxylation of indan by escherichia coli containing the cloned toluene dioxygenase genes from pseudomonas putida f1.escherichia coli jm109(pdtg601), containing the todc1c2ba genes encoding toluene dioxygenase from pseudomonas putida f1, oxidizes indan to (-)-(1r)-indanol (83% r) and trans-1,3-indandiol. under similar conditions, p. putida f39/d oxidizes indan to (-)-(1r)-indanol (96% r), 1-indanone, and trans-1,3-indandiol. the differences in the enantiomeric composition of the 1-indanols formed by the two organisms are due to the presence of a 1-indanol dehydrogenase in p. putida f39/d that preferentially ox ...19921444374
cloning of a creatinase gene from pseudomonas putida in escherichia coli by using an indicator plate.a genomic library of pseudomonas putida dna was constructed by using plasmid pbr322. transformants of escherichia coli in combination with proteus mirabilis cells grown on creatinase test plates were screened for creatinase activity; transformants were considered positive for creatinase activity if a red-pink zone appeared around the colonies. one creatinase-positive clone was further analyzed, and the gene was reduced to a 2.7-kb dna fragment. a unique protein band (with a molecular weight of a ...19921444379
physiological properties of a pseudomonas strain which grows with p-xylene in a two-phase (organic-aqueous) medium.pseudomonas putida idaho utilizes toluene, m-xylene, p-xylene, 1,2,4-trimethylbenzene, and 3-ethyltoluene as growth substrates when these hydrocarbons are provided in a two-phase system at 5 to 50% (vol/vol). growth also occurs on luria-bertani medium in the presence of a wide range of organic solvents. the ability of the organism to grow in the presence of organic solvents is correlated with the logarithm of the octanol-water partition coefficient, with dimethyl-phthalate (log p(oct) = 2.3) bei ...19921444381
effect of pseudobactin 358 production by pseudomonas putida wcs358 on suppression of fusarium wilt of carnations by nonpathogenic fusarium oxysporum fo47.nonpathogenic fusarium oxysporum fo47b10 combined with pseudomonas putida wcs358 efficiently suppressed fusarium wilt of carnations grown in soilless culture. this suppression was significantly higher than that obtained by inoculation of either antagonistic microorganism alone. the increased suppression obtained by fo47b10 combined with wcs358 only occurred when fo47b10 was introduced at a density high enough (at least 10 times higher than that of the pathogen) to be efficient on its own. p. put ...19921444411
genetic variants in the putidaredoxin-cytochrome p-450cam electron-transfer complex: identification of the residue responsible for redox-state-dependent conformers.camphor is hydroxylated in pseudomonas putida by a three-component system comprised of an oxidase, cytochrome p-450cam, and a two-protein electron-transfer chain, putidaredoxin and putidaredoxin reductase [tyson et al. (1972) j. biol. chem. 274, 5777-5784]. the enzymatic removal of putidaredoxin's c-terminal tryptophan is known to cause a much reduced rate of enzymatic activity in the reconstituted camphor hydroxylase system [sligar et al. (1974) proc. natl. acad. sci. u.s.a. 71, 3906-3910]. to ...19921445875
roles of catr and cis,cis-muconate in activation of the catbc operon, which is involved in benzoate degradation in pseudomonas pseudomonas putida, the catbc operon encodes enzymes involved in benzoate degradation. previous studies have determined that these enzymes are induced when p. putida is grown in the presence of benzoate. induction of the enzymes of the catbc operon requires an intermediate of benzoate degradation, cis,cis-muconate, and a regulatory protein, catr. it has been determined that catr binds to a 27-bp region of the catbc promoter in the presence or absence of inducer. we have called this the repres ...19921447146
construction of a broad host range shuttle vector for gene cloning and expression in actinobacillus pleuropneumoniae and other pasteurellaceae.we have constructed a pair of broad host range expression vectors, pjff224-nx and pjff224-xn, based on plasmid rsf1010, which enable cloning and efficient expression of genes in actinobacillus pleuropneumoniae and pasteurella haemolytica and in escherichia coli. the vectors consist of the minimal autonomous replicon of the broad host range plasmid rsf1010 and a type ii chloramphenicol acetyl transferase gene for chloramphenicol resistance selection. in addition, they contain a gene expression ca ...19921448612
dna sequence determination and functional characterization of the oct-plasmid-encoded alkjkl genes of pseudomonas oleovorans.the alkbfghjkl and alkst operons encode enzymes that allow pseudomonas putida (oleovorans) to metabolize alkanes. in this paper we report the nucleotide sequence of a 4592 bp region of the alkbfghjkl operon encoding the alkj, alkk and alkl polypeptides. the alkj gene encodes a protein of 59 kilodaltons. the predicted amino acid sequence shows significant homology with four flavin proteins: choline dehydrogenase, a glucose dehydrogenase and two oxidases. alkj is membrane-bound and converts alipha ...19921453953
[the kinetics of glycol destruction by a pseudomonas putida bs-2 strain].destruction of ethylene glycol and diethylene glycol by pseudomonas putida bs-2 culture under conditions of its batch cultivation has been studied for its physiological regularities. the specific rate of the biomass growth in the region of limiting substrate concentrations depends on the diethylene glycol concentration in the medium and follows the mono equation. a semisaturation constant for diethylene glycol is 209 +/- 17 mg/d. the specific rate of the culture growth is independent of the ethy ...19921453991
p-hydroxyphenylacetate-3-hydroxylase. a two-protein component enzyme.p-hydroxyphenylacetate-3-hydroxylase, an inducible enzyme isolated from the soil bacterium pseudomonas putida, catalyzes the conversion of p-hydroxyphenylacetate to 3,4-dihydroxyphenylacetate. the enzyme requires two protein components: a flavoprotein and a colorless protein referred to as the coupling protein. the flavoprotein alone in the presence of p-hydroxyphenylacetate and substrate analogs catalyzes the wasteful oxidation of nadh with the stoichiometric generation of h2o2. a 1:1 complex o ...19921464599
xyls domain interactions can be deduced from intraallelic dominance in double mutants of pseudomonas putida.the xyls protein is the positive regulator of the tol plasmid-encoded meta-cleavage pathway for the metabolism of alkylbenzoates in pseudomonas putida. this protein is activated by a variety of benzoate analogues. to elucidate the functional domains of the regulator and their interactions, several fusions of the xyls c-terminus to ms2 polymerase and of the n-terminus to beta-galactosidase were constructed but all are inactive. in addition, 15 double mutant xyls genes were constructed in vitro by ...19921465113
expression of the 4-chlorobenzoate dehalogenase genes from pseudomonas sp.-cbs3 in escherichia coli and identification of the gene translation products.the genes encoding the 4-chlorobenzoate dehalogenase of pseudomonas sp. strain cbs3 were, in an earlier study, cloned in escherichia coli dh1 with the cosmid vector ppsa843 and then mobilized to the 4-chlorobenzoate dehalogenase minus strain pseudomonas putida kt2440. in this paper we report on the expression of 4-chlorobenzoate dehalogenase in these clones and on the polypeptide composition of the active enzyme. the dehalogenase activity in whole cells suspended in 3.2 mm 4-chlorobenzoate (30 d ...19921477786
evidence that the hrpb gene encodes a positive regulator of pathogenicity genes from pseudomonas solanacearum.the hrp gene cluster of pseudomonas solanacearum gmi1000 strain encodes functions that are essential for pathogenicity on tomato and for the elicitation of the hypersensitive response on tobacco. in this study, we present the nucleotide sequence of one of the hrp genes (hrpb) located at the left-hand end of the cluster and we show that hrpb encodes a positive regulator controlling the expression of hrp genes. hrpb has a coding capacity for a 477-amino-acid polypeptide, which shows significant si ...19921479894
adaptation of model genetically engineered microorganisms to lake water: growth rate enhancements and plasmid loss.when a genetically engineered microorganism (gem) is released into a natural ecosystem, its survival, and hence its potential environmental impact, depends on its genetic stability and potential for growth under highly oligotrophic conditions. in this study, we compared plasmid stability and potential for growth on low concentrations of organic nutrients of strains of pseudomonas putida serving as model gems. plasmid-free and plasmid-bearing (nah7) prototrophic isogenic strains and two amino-aci ...19921482185
[electron-conformational interactions at the active site of reduced bacterial cytochrome p450cam induced by a substrate and analysis of the electron structure of heme].magnetic circular dichroism (mcd) spectra in the soret region (360-480 nm) of camphor-free and camphor-bound reduced bacterial cytochrome p450cam from pseudomonas putida were recorded and analysed in the temperature range from 2 k to 290 k. the temperature dependences of the mcd intensity are qualitatively changed by binding of substrate to the enzyme. in the absence of camphor the linear increase of the mcd intensity with 1/t at t < 4.2 k gives evidence for degeneracy or near degeneracy of the ...19921491671
nucleotide sequence analysis and comparison of the lexa genes from salmonella typhimurium, erwinia carotovora, pseudomonas aeruginosa and pseudomonas putida.the complete nucleotide sequences of the lexa genes from salmonella typhimurium, erwinia carotovora, pseudomonas aeruginosa and pseudomonas putida were determined; the dna sequences of the lexa genes from these bacteria were 86%, 76%, 61% and 59% similar, respectively, to the escherichia coli k12 gene. the predicted amino acid sequences of the s. typhimurium, e. carotovora and p. putida lexa proteins are 202 residues long whereas that of p. aeruginosa is 204. two putative lexa repressor binding ...19921494343
slow rehydration improves the recovery of dried bacterial populations.slow rehydration of bacteria from dried inoculant formulations provided higher viable counts than did rapid rehydration. estimates were higher when clay and peat powder formulations of rhizobium meliloti, rhizobium leguminosarum biovar trifolii, and pseudomonas putida, with water activities between 0.280 and 0.650, were slowly rehydrated to water activities of approximately 0.992 before continuing the dilution plating sequence. rhizobium meliloti populations averaged 6.8 x 10(8) cfu/g and 1328 c ...19921504917
degradation of the herbicide bromoxynil in pseudomonas putida.biological conversion of the herbicide bromoxynil (3,5-dibromo-4-hydroxybenzonitrile) was studied in a batch culture of pseudomonas putida by using hplc. the process had a cometabolic character and proceeded only in the presence of another, simultaneously metabolizable, carbon and energy source. the intensity of degradation correlated with the growth rate, the degradation stopping when the cosubstrate becomes exhausted or the ph value of the medium falls below 6.5. in a medium with glucose, no l ...19921505868
effect of glucose and ribose on microbial degradation of the herbicide bromoxynil continuously added to soil.bromoxynil (3,5-dibromo-4-hydroxybenzonitrile) was continuously added to chernozem (haplic typic) soil inoculated with a suspension of pseudomonas putida capable of cometabolic decomposition of the compound in a hetero-continuous-flow cultivation setup. in the steady state, when glucose or ribose were simultaneously added, 90 and 47% of the added herbicide was degraded per day, respectively. if the saccharides were absent, only 10-27% of the herbicide was decomposed. addition and removal of gluc ...19921505869
microbial oxidation of adamantanone by pseudomonas putida carrying the camphor catabolic plasmid.intact cells of (+/-)camphor-grown pseudomonas putida, atcc17453(cam), have been shown to oxidize readily the monoketone derivative of cage hydrocarbon adamantane, forming oxygenated products indicative of both biological baeyer-villiger and hydroxylation reactions. formed products were identified as 4-oxahomoadamantan-5-one, 5-hydroxyadamantan-2-one and 1-hydroxy-4-oxahomoadamantan-5-one. minor products formed as a result of secondary reactions were tentatively identified as syn- and anti-1,4-d ...19921510672
lack of homology between two haloacetate dehalogenase genes encoded on a plasmid from moraxella sp. strain b.two genes encoding haloacetate dehalogenases, h-1 and h-2, are closely linked on a plasmid from moraxella sp. strain b. h-1 predominantly acts on fluoroacetate, but h-2 does not. to elucidate the molecular relationship between the two enzymes, we compared their structural genes. two restriction fragments of the plasmid dna were subcloned on m13 phages and their nucleotide sequences were determined. the sequence of each fragment contained an open reading frame that was identified as the structura ...19921512562
construction of a cassette enabling regulated gene expression in the presence of aromatic hydrocarbons.a high-level expression cassette has been constructed from a tol plasmid derived from pseudomonas putida carrying all cis- and trans-acting regulatory elements necessary for transcriptional gene activation in the presence of aromatic hydrocarbons such as toluene. foreign dna can be inserted at unique kpni, saci, and ecori sites 7, 13, and 15 nucleotides downstream of a ribosome binding site. the cassette, flanked by bamhi and ecori restriction sites, was inserted into a broad-host-range vector a ...19921513877
multiple periplasmic catalases in phytopathogenic strains of pseudomonas syringae.phytopathogenic strains of pseudomonas syringae are exposed to plant-produced, detrimental levels of hydrogen peroxide during invasion and colonization of host plant tissue. when p. syringae strains were investigated for their capacity to resist h2o2, they were found to contain 10- to 100-fold-higher levels of total catalase activity than selected strains belonging to nonpathogenic related taxa (pseudomonas fluorescens and pseudomonas putida) or escherichia coli. multiple catalase activities wer ...19921514792
oxidation of nitrotoluenes by toluene dioxygenase: evidence for a monooxygenase reaction.pseudomonas putida f1 and pseudomonas sp. strain js150 initiate toluene degradation by incorporating molecular oxygen into the aromatic nucleus to form cis-1,2-dihydroxy-3-methylcyclohexa-3,5-diene. when toluene-grown cells were incubated with 2- and 3-nitrotoluene, the major products identified were 2- and 3-nitrobenzyl alcohol, respectively. the same cells oxidized 4-nitrotoluene to 2-methyl-5-nitrophenol and 3-methyl-6-nitrocatechol. escherichia coli jm109(pdtg601), which contains the toluene ...19921514810
microbial metabolism of quinoline and related compounds. xiv. purification and properties of 1h-3-hydroxy-4-oxoquinoline oxygenase, a new extradiol cleavage enzyme from pseudomonas putida strain 33/1.1h-3-hydroxy-4-oxoquinoline oxygenase was purified to apparent homogeneity from pseudomonas putida strain 33/1 which can use 1h-4-oxoquinoline as sole source of carbon. the molecular mass of the enzyme was determined to 26,000 da by gel chromatography and by sds polyacrylamide gel electrophoresis. the enzyme is labile at temperatures above 30 degrees c and has a ph optimum of 8.0. it requires oxygen for the reaction and is significantly inhibited by metal ions like cu2+, zn2+, hg2+ and by 4-chlo ...19921515060
a study of the incidence of different fluorescent pseudomonas species and biovars in the microflora of fresh and spoiled meat and fish, raw milk, cheese, soil and water.of 182 various foodstuffs and environmental samples examined, 86% had microflora containing fluorescent pseudomonas in differing proportions. a computer-aided technique was used to identify most of the 445 fluorescent strains. pseudomonas fluorescens biovar v-1 was most frequently isolated (24%); it either predominated or was present in all types of samples. other strains, belonging to the other subgroups of biovar v (v-2, v-4, v-5, v-6 and v-7), together represented 14.3%. we also identified ps ...19921517169
degradation of chlorotoluenes by in vivo constructed hybrid strains: problems of enzyme specificity, induction and prevention of meta-pathway.the activities of the tol plasmid-coded xylene oxygenase, benzylalcohol dehydrogenase, benzaldehyde dehydrogenase of pseudomonas putida strain paw1 were tested with substituted toluenes, benzylalcohols and benzaldehydes, respectively, as substrates. several chlorinated toluenes were shown to induce enzymes of the xylene degradation sequence. conjugative transfer of the tol plasmid from pseudomonas putida strain paw1 to pseudomonas sp. strain b13 and pseudomonas cepacia strain jh230 allowed the i ...19921526468
molecular cloning and characterization of catechol 2,3-dioxygenases from biphenyl/polychlorinated biphenyls-degrading bacteria.catechol 2,3-dioxygenases were cloned from alcaligenes sp. kf711, pseudomonas putida kf715, and achromobacter xylosoxidans kf701 which are biphenyl/polychlorinated biphenyls-degrading bacteria. all of the cloned enzymes were purified by preparative polyacrylamide gel electrophoresis (page). the purified catechol 2,3-dioxygenases were significantly different from one another in ring-fission activities to catechol and its derivatives. the catechol 2,3-dioxygenase from alcaligenes sp. kf711 exhibit ...19921530619
dna homology between siderophore genes from fluorescent pseudomonads.many species of pseudomonads produce fluorescent siderophores involved in iron uptake. we have investigated the dna homology between the siderophore synthesis genes of an opportunist animal pathogen, pseudomonas aeruginosa, and three plant-associated species pseudomonas syringae, pseudomonas putida and pseudomonas sp. b10. there is extensive homology between the dna from the different species, consistent with the suggestion that the different siderophore synthesis genes have evolved from the sam ...19921532617
cytochrome p450cam: crystallography, oxygen activation, and electron transfer.several crystal structures of various substrate and inhibited complexes of the camphor monoxygenase, cytochrome p450cam from pseudomonas putida, are now available. these structures, together with mutagenesis, biochemical, and biophysical studies, have allowed for a detailed penetration into the problem of how p450s activate molecular oxygen, control stereoselectivity, and transfer electrons. this review will provide a summary of the crystallographic work in light of what these structures have ta ...19921537455
sequence and complementation analysis of recf genes from escherichia coli, salmonella typhimurium, pseudomonas putida and bacillus subtilis: evidence for an essential phosphate binding loop.we have compared the recf genes from escherichia coli k-12, salmonella typhimurium, pseudomonas putida, and bacillus subtilis at the dna and amino acid sequence levels. to do this we determined the complete nucleotide sequence of the recf gene from salmonella typhimurium and we completed the nucleotide sequence of recf gene from pseudomonas putida begun by fujita et al. (1). we found that the recf proteins encoded by these two genes contain respectively 92% and 38% amino acid identity with the e ...19921542576
characterization of catechol 2,3-dioxygenases.three catechol 2,3-dioxygenases for biphenyl, naphthalene/salicylate, and toluene/xylene oxidation were cloned from achromobacter xylosoxidans kf701, pseudomonas putida (nah7), and pseudomonas sp. (pwwo). the cloned catechol 2,3-dioxygenases were identified by enzymatic activity assay in addition to yellow bands on polyacrylamide gel after electrophoresis and activity staining. all of the cloned catechol 2,3-dioxygenases exhibited their highest activities on catechol as a substrate compared with ...19921543511
the influence of temperature on the behaviour of mixed bacterial contamination of the shell membrane of the hen's egg.the inner membrane of the air cell of hens' eggs was inoculated with pseudomonas putida, staphylococcus xylosus, enterococcus faecalis, escherichia coli and salmonella enteritidis. the first mentioned eventually dominated the contamination of the albumen of eggs stored at 4, 15, and 20 degrees c. the last mentioned did so in eggs stored at 37 degrees c. the interval between inoculation of the membrane and gross contamination of the albumen was markedly influenced by site of contamination relativ ...19921547832
identification of a cocaine esterase in a strain of pseudomonas maltophilia.a strain of pseudomonas maltophilia (termed mb11l) which was capable of using cocaine as its sole carbon and energy source was isolated by selective enrichment. an inducible esterase catalyzing the hydrolysis of cocaine to ecgonine methyl ester and benzoic acid was identified and purified 22-fold. in the presence of the solubilizing agent cholate, cocaine esterase had a native mr of 110,000 and was shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be a monomer. in the absence ...19921551831
genes and their organization in the replication origin region of the bacterial chromosome.genes and their organization are conserved in the replication origin region of the bacterial chromosome. to determine the extent of the conserved region in gram-positive and gram-negative bacteria, which diverged 1.2 billion years ago, we have further sequenced the region upstream from the dnaa genes in bacillus subtilis and pseudomonas putida. fifteen open reading frames (orfs) and 11 orfs were identified in the 13.6 kb and the 9.8 kb fragments in b. subtilis and p. putida, respectively. eight ...19921552862
cloning and partial sequencing of an operon encoding two pseudomonas putida haloalkanoate dehalogenases of opposite stereospecificity.we have cloned fragments of dna (up to 13 kb), from pseudomonas putida aj1, that code for two stereospecific haloalkanoate dehalogenases. these enzymes are highly specific for d and l substrates. the two genes, designated hadd and hadl, have been isolated and independently expressed in escherichia coli and p. putida hosts by using broad-host-range vectors. they are closely adjacent and inducible in what appears to be an operon with an upstream open reading frame of unknown function. nucleotide s ...19921556080
purification of pro- and eukaryotic superoxide dismutases by charge-controlled hydrophobic chromatography.the process of purifying superoxide dismutases was simplified using charge-controlled hydrophobic chromatography on 10-carboxydecyl sepharose. in only one chromatographic step following ammonium sulphate precipitation, fe-containing superoxide dismutase from pseudomonas putida and cu,zn-containing superoxide dismutase from bovine erythrocytes were purified with an overall yield of about 70% to electrophoretic homogeneity. the specific activities of the crystalline enzyme preparations were expres ...19921560096
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